Purpose: To evaluate the safety and feasibility of topical anesthesia in vitrectomy using the transconjunctival sutureless 25-gauge system (TSV 25G). Methods:TSV 25G vitrectomy was performed prospectively in 46 eyes of 46 patients with macular-based disorders, including macular holes (n = 31), idiopathic epiretinal membranes (n = 11), and vitreoretinal traction syndrome (n = 4). Topical anesthesia was administered in each patient by instilling 2% Alcaine at 5 min intervals, three times before surgery. Additional Alcaine drops were given as a supplementation if needed during the operation. The levels of intraoperative analgesia were graded from 1 (adequate) to 3 (inadequate). Results: Thirty-one patients (67.4%) tolerated the procedure well, with no additional anesthetic agent. Thirteen patients (28.3%) required additional topical anesthesia and 2 patients (4.3%) required a systemic sedative. Intraoperative pain occurred when the sclera was pierced by the microcannula and when sclera indentation was performed. No patient required additional retrobulbar or peribulbar anesthesia. There was no intraoperative complication which was specifically related to topical anesthesia. Conclusion: Topical anesthesia permits the successful management of some surgically less complex vitreoretinal disorders when the TSV 25G is used.